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NA0410

Sigma-Aldrich

GenElute™ HP 无内毒素质粒大提试剂盒

greener alternative

sufficient for 25 preparations

同義詞:

Gen Elute, GenElute 无内毒素质粒试剂盒

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About This Item

分類程式碼代碼:
41105501
NACRES:
NA.52
暫時無法取得訂價和供貨情況

用途

sufficient for 25 preparations

環保替代產品特色

Designing Safer Chemicals
Learn more about the Principles of Green Chemistry.

sustainability

Greener Alternative Product

技術

DNA extraction: suitable

環保替代類別

Aligned,

儲存溫度

15-25°C

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相關類別

一般說明

GenElute HP 无内毒素质粒大量制备试剂盒可以简单、快速地从重组大肠杆菌培养物中分离不含内毒素的质粒DNA。该试剂盒采用了可快速澄清细菌裂解液的滤柱和可捕获质粒DNA的硅胶柱。配套的专利溶液可使质粒DNA结合到结合柱上，同时避免内毒素随之吸附。该技术使得用户可以维持一致的、低于 0.1 EU/μg 质粒DNA的内毒素水平。

制备所得的优质、不含内毒素DNA可直接供包括内毒素敏感细胞转染等最严格的应用使用。

應用

GenElute™ HP Endotoxin-Free Plasmid Maxiprep试剂盒已被用于分离猪肝[1]和大肠杆菌感受态细胞中的质粒。[2]

特點和優勢

  • 可纯化出多达1.2 mg内毒素水平≤0.1 EU/μg高拷贝数质粒DNA
  • 从沉淀细胞到获得纯化的质粒，仅需35分钟
  • 方便的真空形式

原則

通过离心分离法收集过夜的重组大肠杆菌培养液，然后通过改性SDS 碱裂解法进行处理。过滤澄清裂解液，然后加入针对无内毒素质粒制备而优化的结合溶液。随后，质粒捕获到硅胶膜上，同时避免内毒素随之吸附到膜上。通过两遍洗涤步骤，去除污染物。最后，在不含内毒素的水中洗脱结合所得的DNA。回收的质粒DNA主要以超螺旋形式存在。所含的基因组DNA和RNA均低于溴化丙啶染色琼脂糖凝胶电泳的检测水平。

其他說明

更多信息，请见www.sigma-aldrich.com/genelutehp。

法律資訊

GenElute is a trademark of Sigma-Aldrich Co. LLC

訊號詞

Danger

危險分類

Acute Tox. 4 Oral - Eye Irrit. 2 - Flam. Liq. 3 - Met. Corr. 1 - Resp. Sens. 1 - Skin Irrit. 2 - STOT SE 3

標靶器官

Central nervous system

儲存類別代碼

3 - Flammable liquids

閃點（°F）

77.0 °F - closed cup

閃點（°C）

25 °C - closed cup

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分析證明 (COA)

Lot/Batch Number
0000420536
0000420536
0000380779
0000366366
0000380779

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Tan CW, et al.
Parasitology Research, 102(2), 265-275 (2008)
Paul A Beare et al.
Journal of bacteriology, 191(5), 1369-1381 (2008-12-31)
Coxiella burnetii is a gram-negative obligate intracellular bacterium and the causative agent of human Q fever. The lack of methods to genetically manipulate C. burnetii significantly impedes the study of this organism. We describe here the cloning and characterization of
João M Furtado et al.
Investigative ophthalmology & visual science, 53(11), 6856-6862 (2012-09-07)
Toxoplasma gondii, the parasite responsible for ocular toxoplasmosis, accesses the retina from the bloodstream. We investigated the dendritic cell as a potential taxi for T. gondii tachyzoites moving across the human retinal endothelium, and examined the participation of adhesion molecules
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Journal of visualized experiments : JoVE, (64)(64), e3304-e3304 (2012-06-27)
Genome editing is a powerful technique that can be used to elucidate gene function and the genetic basis of disease. Traditional gene editing methods such as chemical-based mutagenesis or random integration of DNA sequences confer indiscriminate genetic changes in an
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相關內容

Endotoxin-free Plasmid DNA Purification

GenElute HP Plasmid Maxiprep Kit isolates endotoxin-free plasmid DNA in 35 minutes, meeting industry standards.

Questions

1–6 of 6 Questions  

  1. Which format gives a better yield, vacuum or spin?

    1 answer

    1. There is no difference in plasmid recovery when using either format.

      Helpful?

  2. Do the binding columns fit on other manufacturers’ manifolds?

    1 answer

    1. Yes. The columns fit on all standard vacuum manifolds.

      Helpful?

  3. What is the expected yield of plasmid DNA that you can isolate from these kits?

    1 answer

    1. Yields can reach as high as 350 mg for a midiprep and 1.2 mg for a maxiprep. As with any protocol, lower yields can be expected with low copy plasmids.

      Helpful?

  4. Is plasmid DNA isolated with this kit suitable for use in transfection?

    1 answer

    1. Yes. However, if you are currently required to use an endofree plasmid purification kit, we recommend using the GenElute HP Endotoxin-free Kit.

      Helpful?

  5. How do the GenElute HP Plasmid Kits differ from other plasmid purification systems?

    1 answer

    1. The GenElute HP Plasmid Kits are much faster than other commercially available kits. A filter syringe is used for preparing the cleared lysate, which replaces a lengthy centrifugation step. These kits also offer the option of using a vacuum or spin purification format rather than a gravity drip to bind, wash and elute the plasmid. In addition, samples are eluted in Tris buffer so there are no alcohol precipitations required.

      Helpful?

  6. How long does it take to isolate plasmid using the vacuum format?

    1 answer

    1. A midiprep or maxiprep requires only 30 minutes to complete.

      Helpful?

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